OEA was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); DSS (molecular weight=36,000–50,000 kDa) was obtained from MP Biochemical (Santa Ana, CA). MPA O-glucuronide and naloxone 3-β-D-glucuronide were obtained from Toronto Research Chemicals Inc. (ON, Canada). β-MCA, UDCA, HDCA, G-LCA, G-UDCA, G-DCA, T-β-MCA, T-UDCA, T-HDCA and T-LCA were purchased from Steraloids Inc. (Newport, RI). α-MCA, CA, DCA, CDCA, lithocholic acid (LCA), G-CDCA, G-CA, T-CDCA, T-DCA, T-CA, dehydrocholic acid and propranolol were purchased from Sigma-Aldrich (St Louis, MO). The anti-UGT1A, anti-FXR, anti-FGF15 and anti-CYP7A1 antibodies were purchased from Santa Cruz Biotechnology. The anti-PPARα antibody was from Abcam (Cambridge, UK). The anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was purchased from SunShine Biotechnology (Nanjing, China). The secondary antibodies and recombinant human FGF19 was obtained from Bioworld Technology (St Louis Park, MN). Other reagents, unless mentioned, were obtained from Sigma-Aldrich. RNA extracts of colon biopsies from 8 healthy humans and 13 ulcerative colitis and Crohn’s disease patients were previously described51 (link). The ulcerative colitis and Crohn’s disease samples were collected from consented patients and approved by the University of Michigan IRB committee (approval number HUM00042210).
Anti pparα antibody
Manufactured by Abcam
Sourced in United States, China, Hong Kong, United Kingdom
Anti-PPARα antibody is a laboratory reagent used in research applications. It is designed to detect the peroxisome proliferator-activated receptor alpha (PPARα) protein, which is a transcription factor that regulates the expression of genes involved in lipid and glucose metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Dehydrocholic acid, by Merck Group (1 mentions)
Gcdca, by Merck Group (1 mentions)
Tcdca, by Merck Group (1 mentions)
G lca, by Steraloids (1 mentions)
G udca, by Steraloids (1 mentions)
G dca, by Steraloids (1 mentions)
T β mca, by Steraloids (1 mentions)
T udca, by Steraloids (1 mentions)
T hdca, by Steraloids (1 mentions)
T lca, by Steraloids (1 mentions)
Lithocholic acid lca, by Merck Group (1 mentions)
Anti cyp7a1, by Santa Cruz Biotechnology (1 mentions)
Anti fxr, by Santa Cruz Biotechnology (1 mentions)
Propranolol, by Merck Group (1 mentions)
Pgc 1α, by Cell Signaling Technology (1 mentions)
Anti ucp1, by Cell Signaling Technology (1 mentions)
Protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Anti perilipin 1, by Cell Signaling Technology (1 mentions)
Anti cpt1, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
5 protocols using anti pparα antibody
1
Inflammatory Bowel Disease Biomarker Analysis
2
Adipose Tissue Protein Analysis
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Samples of epididymal or inguinal white adipose tissues were washed three times with cold PBS before being lysed in radioimmunoprecipitation (RIPA) lysis buffer (10 mmol/L Tris-HCl, pH 7.5, 1% NP-40; 0.1% sodium deoxycholate, 0.2% SDS, 150 mmol/L NaCl, and 1 mmol/L EDTA) supplemented with 1× protease and phosphatase inhibitor cocktail (Thermo, Fremont, CA, USA) on ice. The separated proteins were transferred onto a nitrocellulose membrane. The anti-ATGL (1:100, #2138), anti-HSL (1:200, #4107), anti-p-HSL (1:200. Ser563, #4139), anti-perilipin-1 (1:100, #3467), anti-UCP1 (1:100, #14670), anti-CPT1 (1:100), PGC1α (1:100, #4259), and anti-β-actin (1:3000, #4967) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). The monoclonal anti-PPARα antibody (1:200, ab191226) and secondary antibodies (1:10,000) were obtained from Abcam (Cambridge, MA, USA). Immunoreactive protein bands were visualized using a ChemiDoc XRS+ System (Bio-Rad) and quantified with Gel Pro Analyzer software (Silk Scientific, Inc., Orem, UT, USA). The internal control, β-actin, was used to normalize differences due to loading variations.
3
Molecular Mechanism of m-PEA in NAFLD
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The m-PEA was obtained from Wuxi Cima Science (Wuxi, China). Hematoxylin-Eosin (HE) staining kit was purchased from Solarbio Life Sciences (Beijing, China). Oil Red O was from Sigma-Aldrich (Shanghai, China). Tween-80 and PEG-400 were purchased from Sangon Biotech (Shanghai, China). Anti-PPAR-α antibody (#ab24509) and anti-CD68 antibody (#ab125212) were obtained from Abcam (Shanghai, China). Anti-caspase-1 antibody (clone 14F468) was purchased from Santa Cruz (Shanghai, China). Anti-GAPDH antibody (clone 1E6D9) was purchased from Proteintech (Wuhan, China). Anti-NLRP3 antibody (#A5652) was acquired from ABclonal Technology (Wuhan, China). Anti-α-SMA antibody (clone 1A4) and anti-LC3B antibody (L7543) were obtained from Sigma-Aldrich (Shanghai, China). Antibodies against p62 (#A19700), Beclin1 (#A11761), and ATG7 (#A0691) were obtained from ABconal (Wuhan, China). Horseradish peroxidase (HRP)-conjugated goat anti-mouse and goat anti-rabbit immunoglobulin G (IgG) were purchased from Proteintech (Wuhan, China).
4
Fenofibrate and LPS Regulation of HMGB1 and PPAR-alpha
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Fenofibrate (catalog: F6020) and LPS (catalog: L3755) were purchased from Sigma-Aldrich (St. Louis, MO). All of the cell culture media and supplements were from Sigma. Antibodies against HMGB1 (catalog: 3935) were from Cell Signaling Technology (Danvers, MA). The anti-PPARα antibody (catalog: ab8934) and anti-histone H3 antibody (catalog: ab1791) were obtained from Abcam (Hong Kong). The anti-actin antibody (catalog: A2668) was obtained from Sigma. Primers for quantitative real-time reverse transcriptase-PCR were designed using GenBank sequences.
5
Immunohistochemical and Lipid Staining Protocol
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In preparation for immunohistochemical staining, the 3 µm paraffin sections were dewaxed/deparaffinized and rehydrated. A heat-mediated antigen retrieval procedure in citrate buffer (pH 6.0) was performed. Blocking of endogenous peroxidase was achieved by incubation with 3% H2O2 (Roth, Karlsruhe, Germany) for 10 min at room temperature. Following the blocking with Roti-Block, the sections were incubated with primary antibodies overnight at 4 °C. The following primary antibodies were used: rabbit polyclonal anti-fatty acid synthase (FAS) antibody (Abcam, Camebridge, UK), mouse monoclonal anti-MTCO1 antibody (Abcam, Camebridge, UK) and rabbit polyclonal anti-PPARαantibody (Abcam, Cambridge, UK). After incubation with peroxidase-labeled goat anti-rabbit IgG antibody (KPL, Gaithersburg, MD, USA) or anti-mouse IgG antibody (SeraCare, Milford, MA, USA), di-aminobenzidine (DAB) (DAB-peroxidase substrate kit; Vector Laboratories, Burlingame, CA, USA) was used as a chromogen.
For Oil Red O staining, snap-frozen renal sections (thickness 10 µm) were stained by using Oil Red O solution 0.5% in iso-propanol (Sigma-Aldrich, Merck, Darmstadt, Germany).
For Oil Red O staining, snap-frozen renal sections (thickness 10 µm) were stained by using Oil Red O solution 0.5% in iso-propanol (Sigma-Aldrich, Merck, Darmstadt, Germany).
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